Anti-beta 2 Microglobulin/B2m Antibody

• Catalog Number: orb614096
• Category: Antibodies
• Description: Anti-beta 2 Microglobulin/B2m Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, WB
• Reactivity: Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived rat beta 2 Microglobulin/B2m recombinant protein (Position: I21-M119).
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 13 kDa
• UniProt ID: P01887
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Beta-2-microglobulin; B2m
• Note: For research use only
• Application notes: Western blot, 0.1-0.25μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5μg/ml, Mouse, Rat Flow Cytometry (Fixed), 1-3μg/1x106 cells, Rat ELISA, 0.1-0.5μg/ml,. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of RH35 cells using anti-Beta 2 Microglobulin/B2m antibody. Overlay histogram showing RH35 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Beta 2 Microglobulin/B2m Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Beta 2 Microglobulin/B2m using anti-Beta 2 Microglobulin/B2m antibody. Beta 2 Microglobulin/B2m was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-Beta 2 Microglobulin/B2m Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of Beta 2 Microglobulin/B2m using anti-Beta 2 Microglobulin/B2m antibody. Beta 2 Microglobulin/B2m was detected in immunocytochemical section of NRK cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-Beta 2 Microglobulin/B2m Antibody overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Beta 2 Microglobulin/B2m using anti-Beta 2 Microglobulin/B2m antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: rat thymus tissue lysates, Lane 2: rat lung tissue lysates, Lane 3: rat spleen tissue lysates, Lane 4: rat stomach tissue lysates, Lane 5: rat small intestine tissue lysates, Lane 6: mouse thymus tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Beta 2 Microglobulin/B2m antigen affinity purified polyclonal antibody at 0.25 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Beta 2 Microglobulin/B2m at approximately 13 KD. The expected band size for Beta 2 Microglobulin/B2m is at 13 KD.